Of tumor growth and quantitative RT-PCR was performed (n = 3/group, completed in triplicate; mean SEM shown). (D) Relative expression of genes of p53 signaling, NF-B elements, and c-myc. (E) Heatmap depicting expression of M1/M2 genes. (F) Western blotting was performed for essential members of your p53, NF-B, and MAPK pathways on RAW264.7 cells treated with MQ (blot representative of 2 experiments depicted). P 0.05; P 0.01; P 0.001; P 0.0001 by 2-way ANOVA with many t tests corrected with Bonferroni’s technique (A and D).similarities to those related with the NF-B pathway, a vital mediator of SASP (25) (Supplemental Figure 9E). When comparing the curated set of genes regulated by p53 restoration, the differentially expressed genes in super p53 and APR-246 reated mice revealed outstanding similarities denoted by overlapping expression in those which are up- and downregulated (Supplemental Figure 10A). Further evaluation of immune checkpoint and costimulatory markers in CD4+ and CD8+ T cells in the APR246 reated TME also revealed upregulation of PD-1 and Lag-3 on T cells (Supplemental Figure 10, B and C). Examination of genes related with M1 and M2 polarization inside the non-T cell group also showed decreased expression of M2 genes (Mrc1, Cox2, and Il10) inside the APR-246 reated tumors (Supplemental Figure 10D). We recognized that the sorted non-T cells are heterogeneous and integrated B cells, NK cells, and myeloid cells, which includes TAMs.Adiponectin/Acrp30, Human (277a.a) TAMs can suppress or help T cell proliferation and activity, as well as create cytokines and chemokines that regulate inflammation within the TME which can be vital in antitumor immunity (9). Due to the fact we had currently identified decreased M2 cytokines and chemokines in the tumorlysates, and enhanced T cell proliferation with TAMs from APR-246treated mice, we investigated the transcriptomic consequences of p53-regulated genes in TAMs immediately after APR-246 therapy in sorted TAMs (CD45+CD11b+TCR 4/80+) by quantitative real-time polymerase chain reaction (RT-PCR) (Figure 5D). TAMs from APR-246 reated mice displayed increased Cdkn2a and paradoxical decreases in Mdm2 and p21 (Cdkn1a). Elevated Cdkn2a expression suggests transcriptomic induction of cellular senescence (26). Induction of Cdkn2a expression was lost in TAMs lacking p53 (CSF1R-p53KO). APR-246 therapy was linked with downregulation of NF-B subunit p65 (RelA) and cMyc, both effectors that could handle SASP. Alternatively, TAMs lacking p53 showed dysregulation of NF-B subunits and c-Myc. Additional, TAMs with intact p53 treated with APR246 upregulated Cxcl1, Nos2, and Il12 and downregulated Ccl8 and Mrc1, suggesting transcriptomic suppression of M2 genes and upregulation of M1-associated genes (Figure 5E).SCARB2/LIMP-2, Human (HEK293, His) The transcriptomic reprograming of TAMs induced by APR-246 was lost in mice lacking p53 (CSF1R-p53KO), indicating that the total and efficient TAM polarization from M2 to M1, mediated by APR-246, was dependent on p53.PMID:23776646 J Clin Invest. 2022;132(18):e148141 doi.org/10.1172/JCIThe Journal of Clinical InvestigationWe similarly studied the genes straight regulated by p53 in TAMs from super p53 versus WT mice (Supplemental Figure 8D). Related to that observed in TAMs from APR-246 reated mice, p21 (Cdkn1a) and Mdm2 genes were downregulated, while Cdkn2a was upregulated, suggesting induction of cellular senescence in TAMs from super p53 mice also. In contrast to APR-246 reated TAMs, super p53 erived TAMs had increased p65 (RelA) expression but a comparable decrease in c-Myc. T.
